Bacteria-based gel compositions for topical applications and uses thereof

ABSTRACT

The present invention relates to a composition for use as a medicament, in the form of a gel with characteristics of a reversible thermogel, based on one or more poloxamers, water and optionally excipients, and an effective amount of an active substance. Furthermore, the present invention relates to a process for the preparation of said composition and uses thereof.

The present invention relates to a composition for use as a medicament, in the form of a gel with characteristics of a reversible thermogel, based on one or more poloxamers, water and optionally excipients, and an effective amount of an active substance. Furthermore, the present invention relates to a process for the preparation of said composition and uses thereof.

In the market, there exist various forms of administration of active ingredients, e.g. microorganisms such as bacteria, at least one flavonoid such as rutin, oxerutin, diosmin, hesperidin and troxerutin, chemotherapeutic agents and/or arctigenin, arctin, berberine, berbamine, sanguinarine and chelerythrine, or rutoxides, as such or in the form of plant extracts containing said compounds, or melatonin or derivatives thereof. The topical forms of administration can be patches, creams, gels or suspensions. An example of a gel composition is described in EP2520279 A1.

However, there continues to be a felt need to be able to have a form of administration for topical use for transdermal application that is economical and easy to prepare, stable, efficacious in dosing the effective amount of active substance contained therein, and easily absorbable at the level of the cutis, dermis and epidermis without leaving residues or streaks.

The Applicant has developed a composition in gel form for topical use which provides an adequate and advantageous response to the above-mentioned needs.

The present invention relates to a composition in the form of gel for topical use, having the features as claimed in the accompanying claims.

The present invention relates to a process for the preparation of said composition, having the features as claimed in the accompanying claims.

The present invention relates to a composition for use as a medicament in gel form or topical use, having the features as claimed in the accompanying claims.

The preferred embodiments of the present invention set forth in the description that follows are illustrated solely by way of example and in no way limit the broad scope of application of the present invention, which will appear clear to the person skilled in the art.

In the context of the present invention, composition(s) means a pharmaceutical composition, a composition for medical devices, a composition for dietary supplements or a food composition.

Advantageously, the composition of the present invention is capable of releasing the active substance or active ingredient over time, in a constant, gradual and lasting manner as a controlled transdermal release, in such a way as to prolong its activity and therapeutic effectiveness over time.

Advantageously, the composition of the present invention is a gel in the form of a reversible therrnogel thanks to the presence of a vehicle or carrier which contains specific selected polymers of the poloxamer type.

Advantageously, the composition of the present invention is for topical use for transdermal and/or transmucosal application, said composition is furthermore economical and easy to prepare, stable, efficacious in dosing the effective amount of active substance contained therein over time, and easily absorbable at the level of the cutis, dermis and epidermis without leaving residues or streaks.

The present invention relates to a composition (abbreviated as CMP) in the form of a gel for topical use. Said composition comprises:

-   -   an effective amount of an active substance selected from the         group comprising or, alternatively, consisting of:         (i) at least one microorganism selected from the group         comprising or, alternatively, consisting of: live lactic         bacteria, live bifidobacteria, biologically active bacteria or         active bacterial components, extracts or cell components,         bacterial enzymes, tyndallized bacteria, lysed bacteria,         sonicated bacteria and peptidoglycans; and         (ii) melatonin and/or the natural and/or synthetic derivatives         thereof;         preferably (iii) at least one compound selected from the group         comprising or, alternatively, consisting of a flavonoid such as         rutin, rutoxides, arbutin, oxerutin, diosmin, hesperidin and         troxerutin, chemotherapeutic agents, arctigenin, arctin,         berberine, berbamine, sanguinarine and chelerythrine, as such or         in the form of plant extracts containing said compounds, on         their own or in combination with one or more chemotherapeutic         agents or anti-tumour drugs and a vehicle or carrier, wherein         said composition comprises or, alternatively, consists of an         effective amount of a microorganism (i) selected from the group         comprising or, alternatively, consisting of the microorganisms         in Table 1; preferably said at least one microorganism is         selected from the group comprising or, alternatively, consisting         of the strains: Lactobacillus salivarius (LS01) DSM 22775,         deposited on 23 Jul. 2009; Bifidobacterium breve (BR03) DSM         16604, deposited on 20 Jul. 2004; Lactobacillus pentosus (LPS01)         DSM 21980, deposited on 14 Nov. 2008; Streptococcus thermophilus         (FP4) DSM 18616, deposited on 13 Sep. 2006; Lactobacillus casei         ssp. rhamnosus (LR04) DSM 16605, deposited on 20 Jul. 2004; and         Lactobacillus acidophilus (LA02) DSM 21717, deposited on 6 Aug.         2008; even more preferably the strain is Lactobacillus         salivarius (LS01) DSM 22775, and/or Bifidobacterium breve (BR03)         DSM 16604, and/or L. pentosus (LPS01) DSM 21980, and wherein         said composition is for topical transdermal use in the         preventive and/or curative treatment of pathologies, disorders         or diseases associated with/deriving from alterations of the         immune system selected from the group comprising allergies,         atopy, allergic rhinitis, food hypersensitivity, dermatitis,         atopic dermatitis, eczema, psoriasis, asthma and         immunodeficiencies. Said vehicle or carrier comprises water,         preferably purified water, until reaching 100% by weight of the         composition, and a thickening viscous matrix. Said thickening         viscous matrix comprises or, alternatively, consists of at least         one poloxamer or a mixture of poloxamers and, optionally,         additives and/or excipients and/or adjuvants. Said additives         and/or excipients and/or adjuvants are selected from among those         capable of promoting the formation and stabilisation of the gel         (reversible thermogel) and are selected from the group         comprising or, alternatively, consisting of acid salts, sodium         sorbate, potassium sorbate, sodium benzoate, potassium benzoate,         glycols, ethylene glycol and propylene glycol.

In a preferred embodiment (CMP1), the composition of the present invention (CMP) comprises a poloxamer which is selected from the group comprising or, alternatively, consisting of Poloxamer 124 CAS N. 9003-11-6 with an average molecular weight of 2090-2360, Poloxamer 188 (Lutrol F66) CAS No. 9003-11-6 with an average molecular weight of 7680-9510, Poloxamer 237 CAS N, 9003-11-6 with an average molecular weight of 6840-8830, Poloxamer 338 CAS N 9003-11-6 with an average molecular weight of 12700-17400, Poloxamer 407 (Lutrol® F127 Prill) CAS No. 9003-11-6 with an average molecular weight of 9840-14600, or mixtures thereof; preferably said Poloxamer 188 is present in an amount comprised from 0.1% to 10% by weight, out of 100 g of composition, preferably in an amount comprised from 0.5% to 5% by weight, for example from 1% to 3% by weight; and/or preferably said Poloxamer 407 is present in an amount comprised from 1% to 40% by weight, out of 100 g of composition, preferably in an amount comprised from 5% to 30% by weight, for example from 10% to 20% by weight (composition CMP1). Advantageously, the composition CMP1 contains Poloxamer 188 (Lutrol F66) CAS No. 9003-11-6 with an average molecular weight of 7680-9510 and/or Poloxamer 407 (Lutrol F127 Prill) CAS No. 900-11-6 with an average molecular weight of 984014600.

In another preferred embodiment (CMP4), the composition CMP or CMP1 comprises said microorganisms at a concentration comprised from 1×10⁶ CFU/g to 1×10¹² CFU/g of composition, preferably from 1×10⁷ CFU/g to 1×10¹¹ CFU/g of composition, even more preferably from 1×10⁸ CFU/g to 1×10¹⁰ CFU/g of composition, for example 1×10⁹ CFU/g of composition, and wherein:

-   -   said microorganisms are present in said composition in an amount         by weight comprised from 0.1% to 5%, preferably in an amount by         weight comprised from 0.5% to 3%, even more preferably in an         amount by weight comprised from 1% to 2%, relative to the total         weight of the composition (CMP4).

In another preferred embodiment (CMP5), the composition CMP, CMP1 or 4 comprises an effective amount of an active substance selected from:

(iii) at least one compound selected from the group comprising or, alternatively, consisting of a flavonoid such as rutin, rutoxides, arbutin, oxerutin, diosmin, hesperidin and troxerutin, chemotherapeutic agents, arctigenin, arctin, berberine, berbamine, sanguinarine and chelerythrine, as such or in the form of plant extracts containing said compounds, on their own or in combination with one or more chemotherapeutic agents or anti-tumour drugs; and (ii) melatonin and/or the natural and/or synthetic derivatives thereof; said active substance being present in said composition in an amount by weight comprised from 0.1% to 10% by weight, preferably in an amount comprised from 0.5% to 5% by weight, even more preferably from 1% to 3% by weight, relative to the total weight of the composition (CMP5).

In another embodiment (CMP6), the composition as described above is for use:

-   -   in the treatment of pathologies, disorders or diseases due to         the pathological malfunctioning of physiological functions         connected with circadian rhythms;     -   in the treatment of pathologies, disorders or diseases due to an         abnormal or excessively high or excessively low blood pressure;     -   in the treatment of pathologies, disorders or diseases due to an         excess of free radicals and oxidative stress;     -   in the treatment of pathologies, disorders or diseases due to an         imbalance of the immune system;     -   in the treatment of pathologies, disorders or diseases due to         inflammatory states;     -   in the treatment of pathologies, disorders or diseases due to         acquired immunodeficiency;     -   in the treatment of pathologies, disorders or diseases due to         viral infections and bacterial infections;     -   in a treatment as a support or adjuvant to chemotherapy;     -   in the treatment of cancer with chemotherapeutic agents;     -   in the treatment of sleep disorders in order to favour, in the         treated subjects, the quality of sleep and regularity of sleep,         with particular reference to REM sleep;     -   in the treatment of pathologies, disorders or diseases of the         eyes, of the retina, of the iris, of the optic nerve, retinal         detachment and macular degeneration;     -   in the treatment of vision disorders, in particular in         maculopathy of the retina, via external applications in the         vicinity of the eye and the temple;     -   in the treatment of pathologies, disorders or diseases due to         autism and Down's syndrome, in particular in children and         adolescents, by favouring balance and control also of states of         anxiety;     -   in the treatment of pathologies, disorders or diseases due to         senile dementia or Alzheimer's disease;     -   in the treatment of pathologies, disorders or diseases due to         Parkinson's disease or neurological dysfunctions, by improving         sleep and tremors;     -   in the treatment of pathologies, disorders or diseases due to         pulmonary cystic fibrosis in children, by performing an         anti-inflammatory activity;     -   in the treatment of prostate tumours, by preventing the         propagation thereof;     -   in the treatment of tumours, as an anti-inflammatory,         anti-tumour adjuvant;     -   in the treatment of pathologies, disorders or diseases due to         heart attack, myocardial infarction, cardiopathies, cardiac or         coronary failure or myocardial insufficiency.

The present invention relates to a process for the preparation of a composition in accordance with the compositions CMP or CMP1 or 4 or 5 or 6, said process comprising the following steps:

-   -   mixing in water, under continuous stirring, the various         components, added in succession one after the other or         separately premixed and added together, using a mixer provided         with a temperature control and adjustment means and a stirring         means, wherein the temperature is set at values lower than room         temperature, comprised from 2° C. to 16° C., preferably from         4° C. to 10° C., for example from 6° C. to 8° C., for a time         comprised from 1 minute to 15 minutes so as to obtain a         homogeneous mixture;     -   maintaining said homogeneous mixture, preferably under         continuous stirring, at a temperature comprised from 2° C. to         10° C., preferably at a temperature comprised from 4° C. to 8°         C., for example from 5° C. to 7° C., for a time comprised from 1         minute to 60 minutes, preferably from 5 minutes to 40 minutes,         even more preferably from 10 minutes to 20 minutes, so as to         obtain said composition in a liquid state;     -   packaging said composition in sealed containers.

The vehicle present in the composition of the present invention comprises water and a thickening viscous matrix comprising at least one poloxamer or a mixture of at least due poloxamers.

In the context of the present invention, the term “reversible thermogel” refers to a gel obtained from aqueous copolymer solutions, wherein the copolymer is a poloxamer, capable of gelling in a reversible manner according to the temperature.

Typically, the composition, i.e. the reversible thermogel, of the present invention is liquid at temperatures below room temperature and becomes gelatinous at temperatures close to human body temperature. For example, said composition is liquid at temperatures approximately comprised from 3° C. to 15° C.; in particular, at about 5° C. Said composition in turn gels, that is, becomes a gel, starting from about 20° C. until about 37°; in particular it gels between about 20° and 25° C., preferably, between about 21° and 23° C.

In the context of the present invention, the active agent, for example the melatonin, is present in the composition of the invention in an amount comprised from 0.1% to 3% by weight, out of 100 g of composition; preferably, comprised from 0.5% to 2% by weight, out of 100 g of composition: more preferably, about 1% by weight, out of 100 g of composition; even more preferably, 1% by weight, out of 100 g of composition. The melatonin can be added into the composition as such, in powder, with a purity comprised from 96% to 99.9%; in particular, with a purity comprised from 97% to 99%; for example, with a purity of the 98%. Alternatively, the melatonin can be added into the composition also in the form of microparticles, for example of a size comprised from 50 to 100 μm.

In the context of the present invention, the poloxamers are a series of block copolymers of ethylene oxide and propylene oxide in accordance with the following structural formula;

HO(C₂H₄O)₈(C₃H₆O)_(b)(C₂H₄O)_(a)H,

where a and b are whole numbers expressing the number of the oxyethylene and oxypropylene residues present in the molecule. Poloxamers are non-ionic copolymers of polyoxyethylene-polyoxypropylene used primarily in pharmaceutical formulations as emulsifying, solubilising or welling agents.

The poloxamers of the present invention are capable of thickening the aqueous solutions and giving rise to reversible thermogels possessing rheological properties that vary according to their concentration and molecular weight. A large number of poloxamers (whose molecular weight varies widely according to the values of a and b) can be used for the purposes of the present invention. Among them, the five poloxamers (characterised by the codes 124, 188, 237, 338, 407, respectively) included in the US Pharmacopeia USP NF XVII, in the chapter headed “Poloxamer”, whose contents are incorporated herein in their entirety, have proven to be preferable. Among these, the poloxamers are preferably selected from the group consisting of Poloxamer 188 (Lutrol® F 68, BASF SE, Ludwigshafen, DE), CAS 9003-11-6; Poloxamer 407 (Lutrol® F 127, BASF SE, Ludwigshafen, DE), CAS 900-11-6; or mixtures thereof. In a preferred embodiment of the invention, a mixture of Poloxamer 188 and Poloxamer 407 is used.

In the context of the present invention, the at least one poloxamer or mixture of poloxamers is present in a total amount comprised from 1% to 35% by weight, out of 100 g of composition.

In a preferred embodiment of the invention,

the Poloxamer 188 is present in an amount comprised from 1% to 5% by weight, out of 100 g of composition; preferably, comprised from 1% to 3% by weight, out of 100 g of composition; more preferably, comprised from 1% to 2% by weight, out of 100 g of composition; even more preferably, of 1% by weight, out of 100 g of composition; and the Poloxamer 407 is present in an amount comprised from 15% to 30% by weight, out of 100 g of composition; preferably, comprised from 18% to 25% by weight, out of 100 g of composition; more preferably, comprised from 20% to 23% by weight, out of 100 g of composition; even more preferably, of 21% by weight, out of 100 g of composition.

In the context of the present invention, the water is preferably purified water and is present in an amount such as to reach a balance at 100% by weight of the composition.

The composition of the present invention further comprises known additives/excipients/adjuvants commonly used in the pharmaceutical formulation technique; in particular, said additives/excipients/adjuvants are selected from among those particularly advantageous for the formation and the stabilisation of the reversible thermogel of the invention. In a preferred embodiment of the invention, said additives/excipients/adjuvants are selected from the group consisting of acid salts, such as sodium sorbate, potassium sorbate, sodium benzoate, potassium benzoate; and glycols, such as ethylene glycol and propylene glycol.

In a particularly preferred embodiment, said additives/excipients/adjuvants consist of potassium sorbate, sodium benzoate and propylene glycol, in a total amount comprised from 1 to 5% by weight, out of 100 g of composition.

The pharmaceutical composition for topical use in the form of a reversible thermogel of the present invention can be prepared by adopting well-known apparatus and processing conditions commonly used in the industry for the preparation of a reversible thermogel.

Essentially, the various ingredients making up the desired composition, added in succession or pre-mixed, undergo cold mixing using a suitable mixer provided with a refrigeration temperature control and adjustment means and a stirring means. The mixing temperature is set on values below room temperature, preferably between 3 and 8° C., and the mixture of components is kept under cold stirring for an amount of time sufficient to obtain a completely homogeneous mixture which, in an amount of time that will vary depending on the type of poloxamers used, will transform, still cold, into a completely liquid solution.

The product thus obtained is then packaged in suitable containers, e.g. sealed tubes, and subsequently, on reaching room temperature, will transform into a gel, in particular at the moment when it is applied on the skin.

In one embodiment (FR1), the present invention relates to a composition in the form of a gel for topical use comprising:

-   -   an effective amount of an active substance selected from the         group comprising or, alternatively, consisting of:         (i) at least one microorganism selected from the group         comprising or, alternatively, consisting of: live lactic         bacteria, live bifidobacteria, biologically active bacteria or         active bacterial components, cellular extracts or components,         bacterial enzymes, tyndallized bacteria, lysed bacteria,         sonicated bacteria and peptidoglycans; and/or         (ii) at least one compound selected from the group comprising         or, alternatively, consisting of a flavonoid such as rutin,         rutoxides, arbutin, oxerutin, diosmin, hesperidin and         troxerutin, chemotherapeutic agents, arctigenin, arctin,         berberine, berbamine, sanguinarine and chelerythrine, as such or         in the form of plant extracts containing said compounds, on         their own or in combination with one or more chemotherapeutic         agents or anti-tumour drugs; and/or         (iii) melatonin and/or the natural and/or synthetic derivatives         thereof; and     -   a vehicle or carrier comprising water, a thickening viscous         matrix comprising or, alternatively, consisting of at least one         poloxamer or a mixture of poloxamers and, optionally, additives         and/or excipients and/or adjuvants selected from among those         capable of promoting the formation and stabilisation of the gel,         selected from the group comprising or, alternatively, consisting         of acid salts, sodium sorbate, potassium sorbate, sodium         benzoate, potassium benzoate, glycols, ethylene glycol and         propylene glycol; said composition being for use as a         medicament.

In a preferred embodiment (FR2), the present invention relates to the composition for use according to FR1, wherein said poloxamer is selected from the group comprising or, alternatively, consisting of Poloxamer 124 with an average molecular weight of 2090-2360, Poloxamer 188 (Lutrol F66) CAS No. 9003-11-6 with an average molecular weight of 7680-9510, Poloxamer 237 with an average molecular weight of 6840-8830, Poloxamer 338 with an average molecular weight of 12700-17400, Poloxamer 407 (Lutrol F127 Prill) CAS No. 900-11-6, with an average molecular weight of 984014600, or mixtures thereof; preferably said Poloxamer 188 is present in an amount comprised from 0.1% to 10% by weight, out of 100g of composition, preferably in an amount comprised from 0.5% to 5% by weight, for example from 1% to 3% by weight; and/or preferably said Poloxamer 407 is present in an amount comprised from 1% to 40% by weight, out of 100g of composition, preferably in an amount comprised from 5% to 30% by weight, for example from 10% to 20% by weight.

In a preferred embodiment (FR3), the present invention relates to the composition for use according to FR1, wherein:

-   -   said composition comprises or, alternatively, consists of an         effective amount of an active substance selected from at least         one microorganism (i) and, wherein:     -   said composition is for topical transdermal use in the         preventive and/or curative treatment of pathologies, disorders         or diseases associated with/deriving from alterations of the         immune system selected from the group comprising allergies,         atopy, allergic rhinitis, food hypersensitivity, dermatitis,         atopic dermatitis, eczema, psoriasis, asthma and         immunodeficiencies.

In a preferred embodiment (FR4), the present invention relates to the composition for use according to one of FR1-FR3, wherein said at least one microorganism is selected from the group comprising or, alternatively, consisting of the microorganisms in Table 1; preferably said at least one microorganism is selected from the group comprising or, alternatively, consisting of the strains: Lactobacillus salivarius (LS01) DSM 22775, deposited on 23 Jul 2009; Bifidobacterium breve (BR03) DSM 16604, deposited on 20.072004; Lactobacillus pentosus (LPS01) DSM 21980, deposited on 14 Nov. 2008; Streptococcus thermophilus (FP4) DSM 18616, deposited on 13 Sep. 2006; Lactobacillus casei ssp. rhamnosus (LR04) DSM 16605, deposited on 20 Jul. 2004; and Lactobacillus acidophilus (LA02) DSM 21717, deposited on 6 Aug. 2008; even more preferably the strain is Lactobacillus salivarius (LS01) DSM 22775, and/or Bifidobacterium breve (BR03) DSM 16604, and/or L. pentosus (LPS01) DSM 21980.

In a preferred embodiment (FR5), the present invention relates to the composition for use according to FR4, wherein said microorganisms are present at a concentration comprised from 1×10⁶ CFU/g to 1×10¹² CFU/g of composition, preferably from 1×10⁷ CFU/g to 1×10¹¹ CFU/g of composition, even more preferably from 1×10⁸ CFU/g to 1×10¹⁰ CFU/g of composition, for example 1×10⁹ CFU/g of composition, and wherein:

-   -   said microorganisms are present in said composition in an amount         by weight comprised from 0.1% to 5%, preferably in an amount by         weight comprised from 0.5% to 3%, even more preferably in an         amount by weight comprised from 1% to 2%, relative to the total         weight of composition.

In a preferred embodiment (FR6), the present invention relates to the composition for use according to FR1 or the FR2, wherein:

-   -   said composition comprises or, alternatively, consists of an         effective amount of an active substance selected from:         (ii) at least one compound selected from the group comprising         or, alternatively, consisting of a flavonoid such as rutin,         rutoxides, arbutin, oxerutin, diosmin, hesperidin and         troxerutin, chemotherapeutic agents, arctigenin, arctin,         berberine, berbamine, sanguinarine and chelerythrine, as such or         in the form of plant extracts containing said compounds, on         their own or in combination with one or more chemotherapeutic         agents or anti-tumour drugs; and/or         (iii) melatonin and/or the natural and/or synthetic derivatives         thereof;         said active substance being present in said composition in an         amount by weight comprised from 0.1% to 10% by weight,         preferably in an amount comprised from 0.5% to 5% by weight,         even more preferably from 1% to 3% by weight, relative to the         total weight of the composition.

In a preferred embodiment (FR7), the present invention relates to the composition for use according to one of FR1-FR6, for use:

-   -   in the treatment of pathologies, disorders or diseases due to         the pathological malfunctioning of physiological functions         connected with circadian rhythms;     -   in the treatment of pathologies, disorders or diseases due to an         abnormal or excessively high or excessively low blood pressure;     -   in the treatment of pathologies, disorders or diseases due to an         excess of free radicals and oxidative stress;     -   in the treatment of pathologies, disorders or diseases due to an         imbalance of the immune system;     -   in the treatment of pathologies, disorders or diseases due to         inflammatory states;     -   in the treatment of pathologies, disorders or diseases due to         acquired immunodeficiency;     -   in the treatment of pathologies, disorders or diseases due to         viral infections and bacterial infections;     -   in a treatment as a support or adjuvant to chemotherapy;     -   in the treatment of cancer with chemotherapeutic agents;     -   in the treatment of sleep disorders in order to favour, in the         treated subjects, the quality of sleep and regularity of sleep,         with particular reference to REM sleep;     -   in the treatment of pathologies, disorders or diseases of the         eyes, of the retina, of the iris, of the optic nerve, retinal         detachment and macular degeneration;     -   in the treatment of vision disorders, in particular in         maculopathy of the retina, via external applications in the         vicinity of the eye and the temple;     -   in the treatment of pathologies, disorders or diseases due to         autism and Down's syndrome, in particular in children and         adolescents, by favouring balance and control also of states of         anxiety;     -   in the treatment of pathologies, disorders or diseases due to         senile dementia or Alzheimer's disease;     -   in the treatment of pathologies, disorders or diseases due to         Parkinson's disease or neurological dysfunctions, by improving         sleep and tremors;     -   in the treatment of pathologies, disorders or diseases due to         pulmonary cystic fibrosis in children, by performing an         anti-inflammatory activity;     -   in the treatment of prostate tumours, by preventing the         propagation thereof;     -   in the treatment of tumours, as an anti-inflammatory,         anti-tumour adjuvant;     -   in the treatment of pathologies, disorders or diseases due to         heart attack, myocardial infarction, cardiopathies, cardiac or         coronary failure or myocardial insufficiency,

In a preferred embodiment (FR8), the present invention relates to a process for the preparation of a composition in accordance with any one of FR1-FR7, said process comprising the following steps:

-   -   mixing in water, under continuous stirring, the various         components, added in succession one after the other or         separately premixed and added together, using a mixer provided         with a temperature control and adjustment means and a stirring         means, wherein the temperature is set at values lower than room         temperature, comprised from 2° C. to 16° C., preferably from         4° C. to 10° C., for example from 6° C. to 8° C., for a time         comprised from 1 minute to 15 minutes so as to obtain a         homogeneous mixture;     -   maintaining said homogeneous mixture, preferably under         continuous stirring, at a temperature comprised from 2° C. to         10° C., preferably at a temperature comprised from 4° C. to 8°         C., for example from 5° C. to 7° C., for a time comprised from 1         minute to 60 minutes, preferably from 5 minutes to 40 minutes,         even more preferably from 10 minutes to 20 minutes, so as to         obtain said composition in a liquid state;     -   packaging said composition in sealed containers.

Solely by way of example, which does not limit the various possibilities of production, an example of preparation of a reversible thermogel/composition of the present invention is described here below.

EXAMPLE 1—PREPARATION OF A LOT OF A COMPOSITION (IN THE FORM OF A REVERSIBLE THERMOGEL) OF THE PRESENT INVENTION

Component weight (g) % Mixture of bacteria 2.00 1.00 (1) or (2) or (3) or (4) or (5) Poloxamer 188 4.00 2.00 Poloxamer 407 40.00 20.00 K sorbate 0.40 0.20 Na benzoate 1.00 0.50 Propylene glycol 2.00 1.00 Purified water 150.60 75.30 TOTAL 200.00 100.00

Mixture (1) consisting of only Lactobacillus salivarius (LS01) DSM 22775, or only Bifidobacterium breve (BR03) DSM 16604, or only L. pentosus (LPS01) OSM 21980.

Mixture (2) consisting of Lactobacillus salivarius (LS01) DSM 22775, Bifidobacterium breve (BR03) DSM 16604 and L. pentosus (LPS01) DSM 21980, in a ratio by weight of 1:1:1.

Mixture (3) consisting of Lactobacillus salivarius (LS01) DSM 22775 and Bifidobacterium breve (BR03) DSM 16604 in a ratio by weight of 1:1.

Mixture (4) consisting of Lactobacillus salivarius (LS01) DSM 22775 and L. pentosus (LPS01) OSM 21980, in a ratio by weight of 1:1.

Mixture (5) consisting of Bifidobacterium breve (BR03) DSM 16604 and L. pentosus (LPS01) DSM 21980, in a ratio by weight of 1:1.

The cold purified water (at 5° C.) is weighed into a glass container (for example a beaker) with magnetic stirring, a thermometer, and an external ice bath and the potassium sorbate and sodium benzoate are added under stirring. The mixture is maintained under stirring at 5° C. until complete dissolution of the salts. Then the following are added, again under stirring: the two poloxamers (crystalline), in succession or in a mixture, the methionine (in powder, or in the form of micro-/nanoparticles) and the propylene glycol and cold stirring is continued for at least 15 minutes until a homogeneous appearance is obtained. The container is then placed in a refrigerator at 5° C. for at least 24 h, until obtaining complete dissolution of the ingredients.

After this, the resulting product (liquid) is packaged in 18-20 g tubes and sealed pending use/experimentation. On reaching room temperature and in contact with the skin the product transforms completely into a gel, which is rapidly absorbed following cutaneous and/or transmucosal administration. The pH of the liquid thermogel is 7.40. The gelling temperature is 21°-22° C.

The composition in the form of a thermogel of the present invention, topically administered, in particular on wrists and/or forearms, has shown an excellent cutaneous permeability, triggering a fast onset of the pharmacological action. Furthermore, thanks to the fact of going directly into circulation without passing through the gastrointestinal tract (no metabolism by the liver), it has also shown high blood levels of the active ingredient, for example, melatonin, for a prolonged period of time (slow release). This fact has made it possible to apply low dosages of the drug, thus obtaining the desired advantageous results (modest dosage/high, long-lasting activity) compared to the traditional forms of administration thereof.

For example, a study was conducted on 10 healthy volunteers with different formulations of melatonin, observing the accumulation of melatonin in the saliva over time (it is well known that melatonin spreads passively into saliva via the bloodstream, and its concentration therein represents 24%-33% of its plasma levels, that is to say, the amount of free melatonin not bound to globulins).

The following treatments were administered to the volunteers, randomly divided into 3 groups:

-   -   1. treatment with a formulation of melatonin in a reversible         thermogel in accordance with the present invention (dosage 1         mg/g, single dose)     -   2. treatment with a formulation of melatonin in a reversible         thermogel in accordance with the present invention (dosage 2         mg/g, single dose)     -   3. treatment with a formulation of melatonin in cream (dosage 1         mg/g, single dose)     -   4. treatment with melatonin in tablets (dosage 1 mg/tablet).

A saliva sample was taken at time 0 (basal) and at subsequent times (up to 7 h) after administration, using a Cortisol-Salivette® device (SARSTEDT S.r.l., Verona, IT); the melatonin levels in the saliva were determined by LC-MS (liquid chromatography-mass spectrometry) using a triple quadrupole mass spectrometer (ABSciex QTrap 3200) and substantially following the method of Khan et al. 2013.

The study showed that the formulations of melatonin in a reversible thermogel in accordance with the present invention possess an excellent thermal permeability, a fast onset of the pharmacological action and a lasting duration thereof compared to the other formulations tested.

The pharmaceutical compositions of the present invention also showed to act effectively on all of the patients treated, whereas the commonly used formulations, such as, for example, oral ones, do not act on all individuals.

Furthermore, the pharmaceutical compositions of the present invention have also shown to possess good stability over time.

The pharmaceutical compositions of the present invention are particularly recommended both for transdermal administration and transmucosal administration.

The pharmaceutical compositions of the present invention have shown to be advantageously active, or in any case very promising in the treatment of a number of disorders/diseases due to dysfunctions of physiological functions connected with circadian rhythms such as, for example, the sleep-wake balance; in the regulation of blood pressure; in eliminating of free radicals; in interacting with the immune system, for example, in the treatment of inflammatory states, in the treatment of acquired immunodeficiencies and in the treatment of viral and bacterial infectious diseases and cancer.

Furthermore, the compositions of the present invention have shown to be useful: in the treatment of sleep disorders in all the treated subjects, by favouring the quality of sleep, in particular of REM sleep; in the treatment of the autism and Down syndrome, particularly in children, by favouring their balance; in the treatment of Alzheimer's disease; in the treatment of Parkinson's disease, by improving sleep and tremors: in the treatment of the vision disorders, for example in maculopathy of the retina, via external applications in the vicinity of the eye; in the treatment of the pulmonary manifestations of cystic fibrosis in children, where they provide an anti-inflammatory action; in the treatment of prostate cancer, where they are capable of preventing the propagation thereof; in the treatment of the tumours, as anti-inflammatory, anti-tumour adjuvants.

Commercial Depositary Deposit Deposit No. Name code institution number date Depositor 1 Lactobacillus casei LF1i CNCM I.P. I-785 21 Jul. 1988 Anidral Srl 2 Lactobacillus gasseri LF2i CNCM I.P. I-786 21 Jul. 1988 Anidral Srl 3 Lactobacillus crispatus LF3i CNCM I.P. I-787 21 Jul. 1988 Anidral Srl 4 Lactobacillus fermentum LF4i CNCM I.P. I-788 21 Jul. 1988 Anidral Srl 5 Lactobacillus fermentum LF5 CNCM I.P. I-789 21 Jul. 1988 Anidral Srl 6 Lactobacillus casei ssp. pseudoplantarum LFH i CNCM I.P. I-790 21 Jul. 1988 Anidral Srl 7 Streptococcus thermophilus B39 BCCM LMG LMG P-18383 5 May 1988 Anidral Srl 8 Streptococcus thermophilus T003 BCCM LMG LMG P-18384 5 May 1988 Anidral Srl 9 Lactobacillus pentosus 9/1 ei BCCM LMG LMG P-21019 16 Oct. 2001 Mofin Srl 0 Lactobacillus plantarum 776/1 bi LP 02 BCCM LMG LMG P-21020 16 Oct. 2001 Mofin Srl 11 Lactobacillus plantarum 476LL 20 bi LP 01 BCCM LMG LMG P-21021 16 Oct. 2001 Mofin Srl 12 Lactobacillus plantarum PR ci BCCM LMG LMG P-21022 16 Oct. 2001 Mofin Srl 13 Lactobacillus plantarum 776/2 hi BCCM LMG LMG P-21023 16 Oct. 2001 Mofin Srl 14 Lactobacillus casei ssp. paracasei 181A/3 aiai LPC00 BCCM LMG LMG P-21380 31 Jan. 2002 Anidral Srl 15 Lactobacillus belonging to the acidophilus LA 02 BCCM LMG LMG P-21381 31 Jan. 2002 Anidral Srl group 192A/1 aai 16 Bifidobacterium longum 175A/1 aiai BCCM LMG LMG P-21382 31 Jan. 2002 Anidral Srl 17 Bifidobacterium breve 195A/1 aici BCCM LMG LMG P-21383 31 Jan. 2002 Anidral Srl 18 Bifidobacterium lactis 32A/3 aiai BS 01 BCCM LMG LMG P-21384 31 Jan. 2002 Anidral Srl 19 Lactobacillus plantarum 501/2 gi COAKTIV BCCM LMG LMG P-21385 31 Jan. 2002 Mofin Srl 20 Lactococcus lactis ssp. lactis 504/4 ci BCCM LMG LMG P-21388 31 Jan. 2002 Mofin Srl 21 Lactococcus lactis ssp. lactis 501/4 hi BCCM LMG LMG P-21387 15 Mar. 2002 Mofin Srl 22 Lactococcus lactis ssp. lactis 501/4 ci BCCM LMG LMG P-21388 31 Jan. 2002 Mofin Srl 23 Lactobacillus plantarum 501/4 li BCCM LMG LMG P-21389 15 Mar. 2002 Mofin Srl 24 Lactobacillus acidophilus LA08 BCCM LMG LMG P-26144 3 Nov. 2010 Probiotical SpA 25 Lactobacillus paracasei ssp. paracasei LPC10 BCCM LMG LMG P-26143 3 Nov. 2010 Probiotical SpA 26 Streptococcus thennophilus GB1 DSMZ DSM 16506 18 Jun. 2004 Anidral Srl 27 Streptococcus thermophilus GB5 DSMZ DSM 16507 18 Jun. 2004 Anidral Srl 28 Streptococcus thermophilus Y02 DSMZ DSM 16590 20 Jul. 2004 Anidral Srl 29 Streptococcus thermophilus Y03 DSMZ DSM 16591 20 Jul. 2004 Anidral Srl 30 Streptococcus thermophilus Y04 DSMZ DSM 16592 20 Jul. 2004 Anidral Srl 31 Streptococcus thermophilus YO5 DSMZ DSM 16593 20 Jul. 2004 Anidral Srl 32 = 56 Bifidobacterium adolescentis BA 03 DSMZ DSM 16594 21 Jul. 2004 Anidral Srl 33 Bifidobacterium adolescentis BA 04 DSMZ DSM 16595 21 Jul. 2004 Anidral Srl 34 Bifidobacterium breve BR 04 DSMZ DSM 16596 21 Jul. 2004 Anidral Srl 35 Bifidobacterium pseudocatenulatum BP 01 DSMZ DSM 16597 21 Jul. 2004 Anidral Srl 36 Bifidobacterium pseudocatenulatum BP 02 DSMZ DSM 16598 21 Jul. 2004 Anidral Srl 37 Bifidobacterium longum BL 03 DSMZ DSM 16603 20 Jul. 2004 Anidral Srl 38 Bifidobacterium breve BR 03 DSMZ DSM 16604 20 Jul. 2004 Anidral Srl 39 Lactobacillus casei ssp. rhamnosus LR 04 DSMZ DSM 16605 20 Jul. 2004 Anidral Srl 40 Lactobacillus delbrueckii ssp. bulgaricus LDB 01 DSMZ DSM 16606 20 Jul. 2004 Anidral Srl 41 Lactobacillus delbrueckii ssp. bulgaricus LDB 02 DSMZ DSM 16607 20 Jul. 2004 Anidral Srl 42 Staphylococcus xylosus SX 01 DSMZ DSM 17102 1 Feb. 2005 Anidral Srl 43 = 57 Bifidobacterium adolescentis BA 02 DSMZ DSM 17103 1 Feb. 2005 Anidral Srl 44 Lactobacillus plantarum LP 07 DSMZ DSM 17104 1 Feb. 2005 Anidral Srl 45 Streptococcus thermophilus YO8 DSMZ DSM 17843 21 Dec. 2005 Anidral Srl 46 Streptococcus thermophilus YO9 DSMZ DSM 17844 21 Dec. 2005 Anidral Srl 47 Streptococcus thermophilus YO100 DSMZ DSM 17845 21 Dec. 2005 Anidral Srl 48 Lactobacillus fermentum LF06 DSMZ DSM 18295 24 May 2006 Anidral Srl 49 Lactobacillus fermentum LF07 DSMZ DSM 18296 24 May 2006 Anidral Srl 50 Lactobacillus fermentum LF08 DSMZ DSM 18297 24 May 2006 Anidral Srl 51 Lactobacillus fermentum LF09 DSMZ DSM 18298 24 May 2006 Anidral Srl 52 Lactobacillus gasseri LGS01 DSMZ DSM 18299 24 May 2006 Anidral Srl 53 Lactobacillus gasseri LGA02 DSMZ DSM 18300 24 May 2006 Anidral Srl 54 Lactobacillus gasseri LGS03 DSMZ DSM 18301 24 May 2006 Anidral Srl 55 Lactobacillus gasseri LGS04 DSMZ DSM 18302 24 May 2006 Anidral Srl 56 = 32 Bifidobacterium adolescentis EI-3 BA 03 DSMZ DSM 18350 15 Jun. 2006 Anidral Srl Bifidobacterium catenulatum sp./pseudocatenulatum EI-3I, ID 09-255 57 = 43 Bifidobacterium adolescentis EI-15 BA 02 DSMZ DSM 18351 15 Jun. 2006 Anidral Srl 58 Bifidobacterium adolescentis EI-18 BA 05 DSMZ DSM 18352 15 Jun. 2006 Anidral Srl Bifidobacterium animalis subsp. lacts EI-18, ID 09-256 59 Bifidobacterium catenulatum EI-20 BC 01 DSMZ DSM 18353 15 Jun. 2006 Anidral Srl 60 Streptococcus thermophilus FRai MO1 DSMZ DSM 18613 13 Sep. 2006 Mofin Srl 61 Streptococcus thermophilus LB2bi MO2 DSMZ DSM 18614 13 Sep. 2006 Mofin Srl 62 Streptococcus thermophilus LRci MO3 DSMZ DSM 18615 13 Sep. 2006 Mofin Srl 63 Streptococcus thermophilus FP4 MO4 DSMZ DSM 18616 13 Sep. 2006 Mofin Srl 64 Streptococcus thermophilus ZZ5F8 MO5 DSMZ DSM 18617 13 Sep. 2006 Mofin Srl 65 Streptococcus thermophilus TEO4 MO6 DSMZ DSM 18618 13 Sep. 2006 Mofin Srl 66 Streptococcus thermophilus S1ci MO7 DSMZ DSM 18619 13 Sep. 2006 Mofin Srl 67 Streptococcus thermophilus 641bi MO8 DSMZ DSM 18620 13 Sep. 2006 Mofin Srl 68 Streptococcus thermophilus 277A/1ai MO9 DSMZ DSM 18621 13 Sep. 2006 Mofin Srl 69 Streptococcus thermophilus 277A/2ai MO10 DSMZ DSM 18622 13 Sep. 2006 Mofin Srl 70 Streptococcus thermophilus IDC11 MO11 DSMZ DSM 18623 13 Sep. 2006 Mofin Srl 71 Streptococcus thermophilus ML3di MO14 DSMZ DSM 18624 13 Sep. 2006 Mofin Srl 72 Streptococcus thermophilus TEO3 MO15 DSMZ DSM 18625 13 Sep. 2006 Mofin Srl 73 Streptococcus thermophilus G62 GG1 DSMZ DSM 19057 21 Feb. 2007 Mofin Srl 74 Streptococcus thermophilus G1192 GG2 DSMZ DSM 19058 21 Feb. 2007 Mofin Srl 75 Streptococcus thermophilus GB18 GG3 DSMZ DSM 19059 21 Feb. 2007 Mofin Srl MO2 76 Streptococcus thermophilus CCR21 GG4 DSMZ DSM 19660 21 Feb. 2007 Mofin Srl 77 Streptococcus thermophilus G92 GG5 DSMZ DSM 19061 21 Feb. 2007 Mofin Srl 78 Streptococcus thermophilus G69 GG6 DSMZ DSM 19062 21 Feb. 2007 Mofin Srl 79 Streptococcus thermophilus YO 10 DSMZ DSM 19063 21 Feb. 2007 Anidral Srl 80 Streptococcus thermophilus YO 11 DSMZ DSM 19064 21 Feb. 2007 Anidral Srl 81 Streptococcus thermophilus YO 12 DSMZ DSM 19065 21 Feb. 2007 Anidral Srl 82 Streptococcus thermophilus YO 13 DSMZ DSM 19066 21 Feb. 2007 Anidral Srl 83 Weissella ssp. WSP 01 EX DSMZ DSM 19067 21 Feb. 2007 Anidral Srl 84 Weissella ssp. WSP 02 EX DSMZ DSM 19068 21 Feb. 2007 Anidral Srl 85 Lactobacillus ssp. WSP 03 EX DSMZ DSM 19069 21 Feb. 2007 Anidral Srl 86 Lactobacillus plantarum LP 09 OY DSMZ DSM 19070 21 Feb. 2007 Anidral Srl 87 Lactobacillus plantarum LP 10 OY DSMZ DSM 19071 21 Feb. 2007 Anidral Srl 88 Lactococcus lactis NS 01 DSMZ DSM 19072 21 Feb. 2007 Anidral Srl 89 Lactobacillus fermentum LF 10 DSMZ DSM 19187 20 Mar. 2007 Anidral Srl 90 Lactobacillus fermentum LF 11 DSMZ DSM 19188 20 Mar. 2007 Anidral Srl 91 Lactobacillus casei ssp. rhamnosus LR05 DSMZ DSM 19739 27 Sep. 2007 Anidral Srl 92 Bifidobacterium bifidum BB01 DSMZ DSM 19818 30 Oct. 2007 Anidral Srl 93 Lactobacillus delbrueckii subsp. Lb DSMZ DSM 19948 28 Nov. 2007 Anidral Srl bulgaricus LD 01 94 Lactobacillus delbrueckii subsp. Lb DSMZ DSM 19949 28 Nov. 2007 Anidral Srl bulgaricus LD 02 95 Lactobacillus delbrueckii subsp. Lb DSMZ DSM 19950 28 Nov. 2007 Anidral Srl bulgaricus LD 03 96 Lactobacillus delbrueckii subsp. Lb DSMZ DSM 19951 28 Nov. 2007 Anidral Srl bulgalicus LD 04 97 Lactobacillus delbrueckii subsp. Lb DSMZ DSM 19952 28 Nov. 2007 Anidral Srl bulgaricus LD 05 98 Bifidobacterium pseudocatenulatum B660 DSMZ DSM 21444 13 May 2008 Probiotical SpA 99 Lactobacillus acidophilus LA02 DSMZ DSM 21717 6 Aug. 2008 Probiotical SpA 100 Lactobacillus paracasei LPC 08 DSMZ DSM 21718 6 Aug. 2008 Probiotical SpA 101 Lactobacillus pentosus LPS 01 DSMZ DSM 21980 14 Nov. 2008 Probiotical SpA 102 Lactobacillus rahmnosus LR 06 DSMZ DSM 21981 14 Nov. 2008 Probiotical SpA 103 Lactobacillus delbrueckii ssp. delbrueckii DSMZ 20074 DSMZ DSM 22106 10 Dec. 2008 Probiotical SpA 104 Lactobacillus plantarum LP1 DSMZ DSM 22107 10 Dec. 2008 Probiotical SpA 105 Lactobacillus salivarius LS01 DSMZ DSM 22775 23 Jul. 2009 Probiotical SpA 106 Lactobacillus salivarius LS03 DSMZ DSM 22776 23 Jul. 2009 Probiotical SpA 107 Bifidobacterium bifidum BB01 DSMZ DSM 22892 28 Aug. 2009 Probiotical SpA 108 Bifidobacterium bifidum DSMZ DSM 22893 28 Aug. 2009 Probiotical SpA 109 Bifidobacterium bifidum BB03 DSMZ DSM 22894 28 Aug. 2009 Probiotical SpA 110 Bifidobacterium lactis BS05 DSMZ DSM 23032 13 Oct. 2009 Probiotical SpA 111 Lactobacillus acidophilus LA 06 DSMZ DSM 23033 13 Oct. 2009 Probiotical SpA 112 Lactobacillus brevis LBR01 DSMZ DSM 23034 13 Oct. 2009 Probiotical SpA 113 Bifidobacterium animalis ssp. lactis BS06 DSMZ DSM 23224 12 Jan. 2010 Probiotical SpA 114 Bifidobacterium longum L04 DSMZ DSM 23233 12 Jan. 2010 Probiotical SpA 115 Bifidobacterium longum BL05 DSMZ DSM 23234 12 Jan. 2010 Probiotical SpA 116 Bifidobacterium bifidum MB 109 DSMZ DSM 23731 29 Jun. 2010 Probiotical SpA 117 Bifidobacterium breve MB 113 DSMZ DSM 23732 29 Jun. 2010 Probiotical SpA 118 Bifidobacterium lactis MB 2409 DSMZ DSM 23733 29 Jun. 2010 Probiotical SpA 119 Lactobacillus reuteri LRE01 DSMZ DSM 23877 5 Aug. 2010 Probiotical SpA 120 Lactobacillus reuteri LRE02 DSMZ DSM 23878 5 Aug. 2010 Probiotical SpA 121 Lactobacillus reuteri LRE03 DSMZ DSM 23879 5 Aug. 2010 Probiotical SpA 122 Lactobacillus reuteri LRE04 DSMZ DSM 23880 5 Aug. 2010 Probiotical SpA 123 Lactobacillus paracasei ssp. paracasei LPC09 DSMZ DSM 24243 23 Nov. 2010 Probiotical SpA 124 Lactobacillus acidophilus LA 07 DSMZ DSM 24303 23 Nov. 2010 Probiotical SpA 125 Bifidobacterium bifidum BB04 DSMZ DSM 24437 4 Jan. 2011 Probiotical SpA 126 Lactobacillus crispatus CRL 1251 DSMZ DSM 24438 4 Jan. 2011 Probiotical SpA 127 Lactobacillus crispatus CRL 1266 DSMZ DSM 24439 4 Jan. 2011 Probiotical SpA 128 Lactobacillus paracasei CRL 1289 DSMZ DSM 24440 4 Jan. 2011 Probiotical SpA 129 Lactobacillus Salivarius CRL 1328 DSMZ DSM 24441 4 Jan. 2011 Probiotical SpA 130 Lactobacillus gasseri CRL 1259 DSMZ DSM 24512 25 Jan. 2011 Probiotical SpA 131 Lactobacillus acidophilus CRL 1294 DSMZ DSM 24513 25 Jan. 2011 Probiotical SpA 132 Lactobacillus salivarius LS04 DSMZ DSM 24618 2 Mar. 2011 Probiotical SpA 133 Lactobacillus crispatus LCR01 DSMZ DSM 24619 2 Mar. 2011 Probiotical SpA 134 Lactobacillus crispatus LCR02 DSMZ DSM 24620 2 Mar. 2011 Probiotical SpA 135 Lactobacillus acidophilus LA09 DSMZ DSM 24621 2 Mar. 2011 Probiotical SpA 136 Lactobacillus gasseri LGS05 DSMZ DSM 24622 2 Mar. 2011 Probiotical SpA 137 Lactobacillus paracasei LPC11 DSMZ DSM 24623 2 Mar. 2011 Probiotical SpA 138 Bifidobacterium infantis BI 02 DSMZ DSM 24687 29 Mar. 2011 Probiotical SpA 139 Bifidobacterium bifidum BB 06 DSMZ DSM 24688 29 Mar. 2011 Probiotical SpA 140 Bifidobacterium longum BL 06 DSMZ DSM 24689 29 Mar. 2011 Probiotical SpA 141 Bifidobacterium lactis BS 07 DSMZ DSM 24690 29 Mar. 2011 Probiotical SpA 142 Bifidobacterium longum PCB133 DSMZ DSM 24691 29 Mar. 2011 Probiotical SpA 143 Bifidobacterium breve B632 DSMZ DSM 24706 7 Apr. 2011 Probiotical SpA 144 Bifidobacteriura breve B2274 DSMZ DSM 24707 7 Apr. 2011 Probiotical SpA 145 Bifidobacterium breve B7840 DSMZ DSM 24708 7 Apr. 2011 Probiotical SpA 146 Bifidobacterium longum B1975 DSMZ DSM 24709 7 Apr. 2011 Probiotical SpA 147 Lactobacillus salivarius DLV1 DSMZ DSM 25138 2 Sep. 2011 Probiotical SpA 148 Lactobacillus LRE05 DSMZ DSM 25139 2 Sep. 2011 Probiotical reuteri SpA 149 Lactobacillus LRE06 DSMZ DSM 25140 2 Sep. 2011 Probiotical reuteri SpA 150 Lactobacillus RC 14 DSMZ DSM 25141 2 Sep. 2011 Probiotical reuteri SpA 151 Streptococcus thermophilus ST 10 DSMZ DSM 25246 19 Sep. 2011 Probiotical SpA 152 Streptococcus thermophilus ST 11 DSMZ DSM 25247 19 Sep. 2011 Probiotical SpA 153 Streptococcus thermophilus ST 12 DSMZ DSM 25282 20 Oct. 2011 Probiotical SpA 154 Lactobacillus salivarius DLV8 DSMZ DSM 25545 12 Jan. 2012 Probiotical SpA 155 Bifidobacterium longum DLBL 07 DSMZ DSM 25669 16 Feb. 2012 Probiotical SpA 156 Bifidobacterium longum DLBL 08 DSMZ DSM 25670 16 Feb. 2012 Probiotical SpA 157 Bifidobacterium longum DLBL 09 DSMZ DSM 25671 16 Feb. 2012 Probiotical SpA 158 Bifidobacterium longum DLBL 10 DSMZ DSM 25672 16 Feb. 2012 Probiotical SpA 159 Bifidobacterium longum DLBL 11 DSMZ DSM 25673 16 Feb. 2012 Probiotical SpA 160 Bifidobacterium longum DLBL 12 DSMZ DSM 25674 16 Feb. 2012 Probiotical SpA 161 Bifidobacterium longum DLBL13 DSMZ DSM 25675 16 Feb. 2012 Probiotical SpA 162 Bifidobacterium longum DLBL 14 DSMZ DSM 25676 16 Feb. 2012 Probiotical SpA 163 Bifidobacterium longum DLBL 15 DSMZ DSM 25677 16 Feb. 2012 Probiotical SpA 164 Bifidobacterium longum DLBL 16 DSMZ DSM 25678 16 Feb. 2012 Probiotical SpA 165 Bifidobacterium longum DLBL 17 DSMZ DSM 25679 16 Feb. 2012 Probiotical SpA 166 Lactobacillus johnsonii DLLJO 01 DSMZ DSM 25680 16 Feb. 2012 Probiotical SpA 167 Lactobacillus rhamnosus DLLR 07 DSMZ DSM 25681 16 Feb. 2012 Probiotical SpA 168 Lactobacillus rhamnosus DLLR 08 DSMZ DSM 25632 16 Feb. 2012 Probiotical SpA 169 Lactobacillus reuteri DLLRE 07 DSMZ DSM 25683 16 Feb. 2012 Probiotical SpA 170 Lactobacillus reuteri DLLRE 08 DSMZ DSM 25684 16 Feb. 2012 Probiotical SpA 171 Lactobacillus reuteri DLLRE 09 DSMZ DSM 25685 16 Feb. 2012 Probiotical SpA 172 Bifidobacterium longum DLBL 18 DSMZ DSM 25708 24 Feb. 2012 Probiotical SpA 173 Bifidobacterium infantis BI 03 DSMZ DSM 25709 24 Feb. 2012 Probiotical SpA 174 Lactobacillus plantarum LP 09 DSMZ DSM 25710 24 Feb. 2012 Probiotical SpA 175 Bifidobacterium longum DLBL 19 DSMZ DSM 25717 1 Mar. 2012 Probiotical SpA 176 Bifidobacterium longum DLBL 20 DSMZ DSM 25718 1 Mar. 2012 Probiotical SpA 177 Lactobacillus salivarius LS 05 DSMZ DSM 26036 6 Jun. 2012 Probiotical SpA 178 Lactobacillus salivatius LS 06 DSMZ DSM 26037 6 Jun. 2012 Probiotical SpA 179 Lactobacillus pentosus LPS 02 DSMZ DSM 26038 6 Jun. 2012 Probiotical SpA 180 Bifidobacterium pseudolongum BPS 01 DSMZ DSM 26456 2 Oct. 2012 Probiotical ssp. globosum SpA 181 Lactobacillus fermentum LF15 DSMZ DSM 26955 1 Mar. 2013 Probiotical SpA 182 Lactobacillus fermentum LF16 DSMZ DSM 26956 1 Mar. 2013 Probiotical SpA 183 Lactobacillus casei LC03 DSMZ DSM 27537 24 Jul. 2013 Probiotical SpA 184 Lactobacillus crispatus LCR03 DSMZ DSM 27538 24 Jul. 2013 Probiotical SpA 185 Lactobacillus jensenii LJE01 DSMZ DSM 27539 24 Jul. 2013 Probiotical SpA 186 Lactobacillus helveticus ID 922 LH01 DSMZ DSM 28153 4 Dec. 2013 Probiotical SpA 187 Lactobacillus helveticus ID 923 LH02 DSMZ DSM 28154 4 Dec. 2013 Probiotical SpA 188 Lactococcus lactis ssp. cremoris ID 1612 LLC02 DSMZ DSM 28155 4 Dec. 2013 Probiotical SpA 189 Lactococcus lactis ssp. cremoris ID 1252 LLC03 DSMZ DSM 28156 4 Dec. 2013 Probiotical SpA 190 Lactococcus lactis ssp. Lactis ID 1254 LLL01 DSMZ DSM 28157 4 Dec. 2013 Probiotical SpA 191 Bifidobacterium longum BL 01 DSMZ DSM 28173 11 Dec. 2013 Probiotical SpA 192 Bifidobacterium lungum BL 02 DSMZ DSM 28174 11 Dec. 2013 Probiotical SpA 193 Bifidobaterium animalis ssp. lactis Bb1 DSMZ DSM 17850 23 Dec. 2005 BiaMan Srl 194 Streptococcus thermophilus ST 16 BM DSMZ DSM 19526 13 Jul. 2007 BioMan Srl 195 Bifidobacterium infantis BI 04 DSMZ DSM 28651 8 Apr. 2014 Probiotical SpA 196 Bifidobacterium infantis BI 05 DSMZ DSM 28652 8 Apr. 2014 Probiotical SpA 197 Streptococcus thermophilus ST 15 DSMZ DSM 28911 11 Jun. 2014 Probiotical SpA 198 Streptococcus thermophilus ST 16 DSMZ DSM 28912 11 Jun. 2014 Probiotical SpA 199 Streptococcus thermophilus ST 17 DSMZ DSM 28913 11 Jun. 2014 Probiotical SpA 200 Lactobacillus fermentum LF18 DSMZ DSM 29197 30 Jul. 2014 Probiotical SpA 201 Lactobacillus fermentum LF19 DSMZ DSM 29198 30 Jul. 2014 Probiotical SpA 202 Leuconostoc sp. LM01 DSMZ DSM 29372 10 Sep. 2014 Mofin Srl 203 Leuccnostoc sp. LM10 DSMZ DSM 29373 10 Sep. 2014 Mofin Srl 204 Leuconostoc sp. LM11 DSMZ DSM 29374 10 Sep. 2014 Mofin Srl 205 Leuconostoc sp. LM12 DSMZ DSM 29375 10 Sep. 2014 Mofin Srl 206 Lactobacillusplantarum LP10 DSMZ DSM 29389 10 Sep. 2014 Mofin Srl 207 Lactobacillusplantarum LP11 DSMZ DSM 29390 10 Sep. 2014 Mofin Srl 208 Lactobacillusplantarum LP12 DSMZ DSM 29400 10 Sep. 2014 Mofin Srl 209 Lactobacillusplantarum LP13 DSMZ DSM 29401 10 Sep. 2014 Mofin Srl 210 Lactobacillus pentosus LPS03 DSMZ DSM 29402 10 Sep. 2014 Mofin Srl 211 Lactobacillus reuteri LRE10 DSMZ DSM 29403 10 Sep. 2014 Mofin Srl 212 Lactobacillus brevis LBR02 DSMZ DSM 29404 10 Sep. 2014 Mofin Srl 213 Lactobacillus salivarius LS 07 DSMZ DSM 29476 9 Oct. 2014 Probiotical SpA 214 Bifidobacterium breve BR 05 DSMZ DSM 29494 9 Oct. 2014 Probiotical SpA 215 Lactococcus lactis ssp. cremoris LCC02 DSMZ DSM 29536 22 Oct. 2014 Probiotical SpA 216 Bifidobacterium longum BL 21 DSMZ DSM 29884 15 Jan. 2015 Probiotical SpA 217 Lactobacillus rhamnosus LR 09 DSMZ DSM 29885 15 Jan. 2015 Probiotical SpA 218 Lactobacillus kefiri LKE01 DSMZ DSM 32027 8 Apr. 2015 Probiotical SpA 219 Lactobacillus kefiri LKE02 DSMZ DSM 32056 29 May 2015 Probiotical SpA 220 Lactobacillus acidophilus LA10 DSMZ DSM 32075 3 Jul. 2015 Probiotical SpA 221 Lactobacillus kefiranofaciens LKR01 DSMZ DSM 32076 3 Jul. 2015 Probiotical SpA 222 Lactobacillus kefiri LKF01 DSMZ DSM 32079 10 Jul. 2015 Probiotical SpA 223 Lactobaciullus kefiri LKF02 DSMZ DSM 32080 10 Jul. 2015 Probiotical SpA 224 Streptococcus thermophilus ST18 DSMZ DSM 32134 3 Sep. 2015 Mofin S.r.l. 225 Streptococcus thermophilus ST19 DSMZ DSM 32135 3 Sep. 2015 Mofin S.r.l. 226 Streptococcus thermophilus ST20 DSMZ DSM 32136 3 Sep. 2015 Mofin S.r.l. 227 Streptococcus thermophilus ST21 DSMZ DSM 32137 3 Sep. 2015 Mofin S.r.l. 228 Streptococcus thermophilus ST22 DSMZ DSM 32138 3 Sep. 2015 Mofin S.r.l. 229 Streptococcus thermophilus ST23 DSMZ DSM 32139 3 Sep. 2015 Mofin S.r.l. 230 Streptococcus thermophilus ST24 DSMZ DSM 32140 3 Sep. 2015 Mofin S.r.l. 231 Lactobacillus salivarius LS02 DSMZ DSM 32204 13 Nov. 2015 Probiotical SpA 232 Weissella confusa WC01 DSMZ DSM 32156 22 Sep. 2015 Mofin S.r.l. 233 Weissella confusa WC02 DSMZ DSM 32157 22 Sep. 2015 Mofin S.r.l. 234 Lactobacillus curvatus LCU01 DSMZ DSM 32160 22 Sep. 2015 Mofin S.r.l. 235 Lactobacillus plantarum LMC1 DSMZ DSM 32252 29 Sep. 2016 Probiotical SpA 236 Lactobacillus reuteri LMC3 DSMZ DSM 32253 29 Sep. 2016 Probiotical SpA 237 Lactobacillus parasei LMC4 DSMZ DSM 32254 29 Sep. 2016 Probiotical SpA 238 Lactobacillus reuteri LMC5 DSMZ DSM 32255 29 Sep. 2016 Probiotical SpA 239 Lactobacillus rhamnosus LMC6 DSMZ DSM 32256 29 Sep. 2016 Probiotical SpA 240 Lactobacillus rhamnosus LMC7 DSMZ DSM 32257 29 Sep. 2016 Probiotical SpA 241 Lactobacillus paracasei LMC8 DSMZ DSM 32258 29 Sep. 2016 Probiotical SpA 242 Lactobacillus reuteri LMC9 DSMZ DSM 32259 29 Sep. 2016 Probiotical SpA 243 Lactobacillus rhamnosus LMC10 DSMZ DSM 32260 29 Sep. 2016 Probiotical SpA 244 Lactobacillus fermentum LF25 DSMZ DSM 32275 15 Mar. 2016 Probiotical SpA 245 Lactobacillus fermentum LF5 DSMZ DSM 32277 18 Mar. 2016 Probiotical SpA 246 Lactobacillus fermentum LF20 DSMZ DSM 32288 14 Apr. 2016 Probiotical SpA

Materials and Methods Experimental Protocol

The experiment was conducted on a group of 6 volunteers, all female and aged between 29 and 41 years (mean age 34.5 years).

Each volunteer tested 5 formulations of melatonin (gel 0.1%, gel 0.2%, gel 1%, cream, Circadin 2 mg), with a wash-out period of one week after each application.

The saliva samples were taken according to the following experimental scheme:

h 10.00 Basal

h 10.30 30′ h 11.00 1 h h 13.00 3 h h 16.00 6 h

The saliva was collected with Salivette (Sarstedt), following the instructions attached thereto (mouth rinsing before each sampling, chewing on the swab for 1 minute). The saliva was extracted from the swab by 2 min centrifugation at 4000 rpm, then divided into 300 μl aliquots and frozen at −20° C. until the time of analysis.

Data Analysis

The melatonin assays were performed with the ELISA (Enzyme-Linked Immunosorbent Assay) technique, using the Melatonin (direct) Saliva kit (SLV-4779-DRG Instruments, Germany). The analytical sensitivity and inter- and intra-assay coefficients of variation are the following: 0.3 pg/ml, 7.6-13.0% and 6.1-10.8%.

The kit in question is certified for in-vitro diagnostic use.

Patient ID Gel 0.1% 1 mg/g 1 0.42 6.65 7.91 18.2 14.6 37.76 2 0.33 4.88 4.22 4.72 2.33 16.48 3 0.1 1.6 1.62 7.55 2.09 10.94 4 1.93 17.9 22.4 18.7 9.46 70.39 5 0.1 5.67 9.59 2.43 1.67 19.46 6 0.14 14.4 8.35 10.10 8.70 71.56 Basal 30′ 1 h 3 h 6 h total Total 47.87 226.04 mean = 37.67 

1. A composition in the form of a gel for topical use comprising: an effective amount of an active substance selected from the group comprising or, alternatively, consisting of: (i) at least one microorganism selected from the group comprising or, alternatively, consisting of: live lactic bacteria; live bifidobacteria, biologically active bacteria or active bacterial components, extracts or cell components, bacterial enzymes, tyndallized bacteria, lysed bacteria, sonicated bacteria and peptidoglycans; and (ii) melatonin and/or the natural and/or synthetic derivatives thereof; and a vehicle or carrier comprising water, a thickening viscous matrix comprising or, alternatively, consisting of at least one poloxamer or a mixture of poloxamers and, optionally, additives and/or excipients and/or adjuvants selected from among those capable of promoting the formation and stabilisation of the gel, selected from the group comprising or, alternatively, consisting of acid salts, sodium sorbate, potassium sorbate, sodium benzoate, potassium benzoate, glycols, ethylene glycol and propylene glycol; wherein said composition comprises or, alternatively, consists of an effective amount of a microorganism (i) selected from the group comprising or, alternatively, consisting of the microorganisms in Table 1; preferably said at least one microorganism is selected from the group comprising or, alternatively, consisting of the strains: Lactobacillus saliverius (LS01) DSM 22775, deposited on 23 Jul. 2009; Bifidobacterium breve (BR03) DSM 16604, deposited on 20 Jul. 2004; Lactobacillus pentosus (LPS01) DSM 21980, deposited on 14 Nov. 2008; Streptococcus thermophilus (FP4) DSM 18616, deposited on 13 Sep. 2006; Lactobacillus casei ssp. rhamnosus (LR04) DSM 16605, deposited on 20 Jul. 2004; and Lactobacillus acidophilus (LA02) DSM 21717, deposited on 6 Aug. 2008; even more preferably the strain is Lactobacillus salivarius (LS01) DSM 22775, and/or Bifidobacterium breve (BR03) DSM 16604, and/or L. pentosus (LPS01) DSM 21980, and wherein said composition is for topical transdermal use in the preventive and/or curative treatment of pathologies, disorders or diseases associated with/deriving from alterations of the immune system selected from the group comprising allergies, atopy, allergic rhinitis, food hypersensitivity, dermatitis, atopic dermatitis, eczema, psoriasis, asthma and immunodeficiencies.
 2. The composition for use according to claim 1, wherein said poloxamer is selected from the group comprising or, alternatively, consisting of Poloxamer 124 CAS N. 9003-11-6 with an average molecular weight of 2090-2360, Poloxamer 188 (Lutrol® F66) CAS No. 9003-11-6 with an average molecular weight of 7680-9510, Poloxamer 237 with an average molecular weight of 6840-8830, Poloxamer 338 with an average molecular weight of 12700-17400, Poloxamer 407 (Lutrol® F127 Prill) CAS No. 9003-11-6 with an average molecular weight of 9840-14600, or mixtures thereof; preferably said Poloxamer 188 is present in an amount comprised from 0.1% to 10% by weight, out of 100 g of composition, preferably in an amount comprised from 0.5% to 5% by weight, for example from 1% to 3% by weight; and/or preferably said Poloxamer 407 is present in an amount comprised from 1% to 40% by weight, out of 100 g of composition, preferably in an amount comprised from 5% to 30% by weight, for example from 10% to 20% by weight.
 3. The composition for use in accordance with one of claims 1-2, further comprising, as an active substance (iii), an effective amount of at least one compound selected from the group comprising or, alternatively, consisting of a flavonoid such as rutin, rutoxides, arbutin, oxerutin, diosmin, hesperidin and troxerutin, chemotherapeutic agents, arctigenin, arctin, berberine, berbamine, sanguinarine and chelerythrine, as such or in the form of plant extracts containing said compounds, on their own or in combination with one or more chemotherapeutic agents or anti-tumour drugs.
 4. The composition for use in accordance with any one of the preceding claims, wherein: said microorganisms are present at a concentration comprised from 1×10⁶ CFU/g to 1×10¹² CFU/g of composition, preferably from 1×10⁷ CFU/g to 1×10¹¹ CFU/g of composition, even more preferably from 1×10⁸ CFU/g to 1×10¹⁰ CFU/g of composition, for example 1×10⁹ CFU/g of composition, and wherein: said microorganisms are present in said composition in an amount by weight comprised from 0.1% to 5%, preferably in an amount by weight comprised from 0.5% to 3%, even more preferably in an amount by weight comprised from 1% to 2%, relative to the total weight of composition.
 5. The composition for use in accordance with claim 1 or 2, wherein: said composition comprises an effective amount of an active substance selected from: (iii) at least one compound selected from the group comprising or, alternatively, consisting of a flavonoid such as rutin, rutoxides, arbutin, oxerutin, diosmin, hesperidin and troxerutin, chemotherapeutic agents, arctigenin, arctin, berberine, berbamine, sanguinarine and chelerythrine, as such or in the form of plant extracts containing said compounds, on their own or in combination with one or more chemotherapeutic agents or anti-tumour drugs; and/or (ii) melatonin and/or the natural and/or synthetic derivatives thereof; said active substance being present in said composition in an amount by weight comprised from 0.1% to 10% by weight, preferably in an amount comprised from 0.5% to 5% by weight, even more preferably from 1% to 3% by weight, relative to the total weight of the composition.
 6. The composition in accordance with any one of claims 1-5, for use: in the treatment of pathologies, disorders or diseases due to the pathological malfunctioning of physiological functions connected with circadian rhythms; in the treatment of pathologies, disorders or diseases due to an abnormal or excessively high or excessively low blood pressure; in the treatment of pathologies, disorders or diseases due to an excess of free radicals and oxidative stress; in the treatment of pathologies, disorders or diseases due to an imbalance of the immune system; in the treatment of pathologies, disorders or diseases due to inflammatory states; in the treatment of pathologies, disorders or diseases due to acquired immunodeficiency; in the treatment of pathologies, disorders or diseases due to viral infections and bacterial infections; in a treatment as a support or adjuvant to chemotherapy; in the treatment of cancer with chemotherapeutic agents; in the treatment of sleep disorders in order to favour, in the treated subjects, the quality of sleep and regularity of sleep, with particular reference to REM sleep; in the treatment of pathologies, disorders or diseases of the eyes, of the retina, of the iris, of the optic nerve, retinal detachment and macular degeneration; in the treatment of vision disorders, in particular in maculopathy of the retina, via external applications in the vicinity of the eye and the temple; in the treatment of pathologies, disorders or diseases due to autism and Down's syndrome, in particular in children and adolescents, by favouring balance and control also of states of anxiety; in the treatment of pathologies, disorders or diseases due to senile dementia or Alzheimer's disease; in the treatment of pathologies, disorders or diseases due to Parkinson's disease or neurological dysfunctions, by improving sleep and tremors; in the treatment of pathologies, disorders or diseases due to pulmonary cystic fibrosis in children, by performing an anti-inflammatory activity; in the treatment of prostate tumours, by preventing the propagation thereof; in the treatment of tumours, as an anti-inflammatory, anti-tumour adjuvant; in the treatment of pathologies, disorders or diseases due to heart attack, myocardial infarction, cardiopathies, cardiac or coronary failure or myocardial insufficiency.
 7. A process for the preparation of a composition in accordance with any one of the preceding claims, said process comprising the following steps: mixing in water, under continuous stirring, the various components, added in succession one after the other or separately premixed and added together, using a mixer provided with a temperature control and adjustment means and a stirring means, wherein the temperature is set at values lower than room temperature, comprised from 2° C. to 16° C., preferably from 4° C. to 10° C., for example from 6° C. to 8° C., for a time comprised from 1 minute to 15 minutes so as to obtain a homogeneous mixture; maintaining said homogeneous mixture, preferably under continuous stirring, at a temperature comprised from 2° C. to 10° C., preferably at a temperature comprised from 4° C. to 8° C., for example from 5° C. to 7° C., for a time comprised from 1 minute to 60 minutes, preferably from 5 minutes to 40 minutes, even more preferably from 10 minutes to 20 minutes, so as to obtain said composition in a liquid state; packaging said composition in sealed containers. 